LOCUS       sequence-349833-        7506 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pGBKT7 yeast two-hybrid plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7506)
  AUTHORS   Bentham AR, Petit-Houdenot Y, Win J, Chuma I, Terauchi R, Banfield 
            MJ, Kamoun S, Langner T
  TITLE     A single amino acid polymorphism in a conserved effector of the 
            multihost blast fungus pathogen expands host-target binding 
            spectrum.
  JOURNAL   PLoS Pathog. 2021 Nov 10;17(11):e1009957. doi: 
            10.1371/journal.ppat.1009957. eCollection 2021 Nov.
  PUBMED    34758051
REFERENCE   2  (bases 1 to 7506)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2021-11-10"; volume: "17(11):e1009957. doi"; pages:
            " 10.1371/journal.ppat.1009957. eCollection 2021 Nov"
COMMENT     Sequence Label: pTL382
FEATURES             Location/Qualifiers
     source          1..7506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     1213..1232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      1541..1588
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1615..1802
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1670..1694)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1826..1842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1826..1842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1839..1861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1850..1866
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1874..1880,1881..1898,1899..1904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1919..1940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2057..2074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2228..2816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2308..2327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3145..3192
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3217..3236
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3424..4218)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3424..4218)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3535..3554)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4145..4164
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4219..4323)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4350..5692
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5313..5360)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     5733..5751
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5789..5811)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5911..5930
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5951..6232
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6233..6907
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7010..7465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7147..7168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7359..7378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcatggcc gtggtggaac
     1321 tgaaagtggg tatgcactgc gatcgctgca tcaaggccat taaaaaagcc attaagacca
     1381 ttgacgatat ggagagctac cagctggaaa ctgagatcaa caaagtgacc gtgaccggta
     1441 acgtgacccc ggatgaagtg gtgaaagccc tgcagaagat cggcaaaacc gccaccaatt
     1501 ggggtgaaga tggatccgtc gacctgcagc ggccgcataa ctagcataac cccttggggc
     1561 ctctaaacgg gtcttgaggg gttttttgcg cgcttgcagc caagctaatt ccgggcgaat
     1621 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa
     1681 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg
     1741 taggtcaggt tgctttctca ggtatagcat gaggtcgctc ttattgacca cacctctacc
     1801 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1861 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1921 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1981 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2041 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2101 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2161 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2221 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2281 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2341 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2401 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2461 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2521 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2581 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2641 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2701 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2761 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2821 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2881 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2941 gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc
     3001 cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg
     3061 aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc
     3121 cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc
     3181 tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta
     3241 gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     3301 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     3361 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     3421 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     3481 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     3541 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     3601 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgagt
     3661 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     3721 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     3781 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     3841 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     3901 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     3961 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     4021 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     4081 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     4141 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     4201 tccatcttgt tcaatcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4321 gcgcacattt ccccgaaaag tgccacctga acgaagcatc tgtgcttcat tttgtagaac
     4381 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     4441 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt
     4501 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     4561 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     4621 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     4681 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     4741 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     4801 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     4861 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     4921 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     4981 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     5041 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     5101 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     5161 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     5221 tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc
     5281 ttcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct agagaatagg
     5341 aacttcggaa taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc
     5401 gagctgcgca catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata
     5461 tatatataca tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc
     5521 gtctatttat gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg
     5581 cggggtatcg tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct
     5641 caattggatt agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcatactaag
     5701 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5881 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5941 accatagatc aacgacatta ctatatatat aatataggaa gcatttaata gaacagcatc
     6001 gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag atattcttta
     6061 ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct ttttttgccg
     6121 attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg
     6181 actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt
     6241 tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga
     6301 ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat gtgtgcccaa
     6361 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata
     6421 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga
     6481 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga
     6541 gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc
     6601 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt
     6661 gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg
     6721 ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg gactgacgcc
     6781 agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg
     6841 tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa
     6901 gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt gccgatctat
     6961 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     7021 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     7081 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     7141 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     7201 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     7261 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     7321 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     7381 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     7441 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     7501 ggcgat
//
